CSIRO PUBLISHING Australian Journal of Botany, 2021, 69, 113–120 https://doi.org/10.1071/BT20122 The contribution of pathogenic soil microbes to ring formation in an iconic Australian arid grass, Triodia basedowii (Poaceae) Neil D. Ross A and Angela T. Moles A,B AEvolution & Ecology Research Centre, School of Biological, Earth and Environmental Sciences, UNSW Sydney, NSW 2052, Australia. BCorresponding author. Email: [email protected] Abstract. Ring-forming species of spinifex grasses (Triodia spp.) are a dominant feature across much of Australia’s arid and semi-arid zone. Researchers have long been curious about the mechanisms underpinning their striking growth form. However, none of the factors investigated to date provide a convincing explanation for ring formation. Here, we asked whether an accumulation of pathogenic soil microbes might impede seedling emergence and subsequent growth in the centre of Triodia basedowii rings. We collected soil from inside and outside naturally occurring spinifex rings and compared plants grown in soil with live microbes to plants grown in sterilised soil. Consistent with our hypothesis, we found that emergence of T. basedowii seedlings was lower in live soil from inside the rings than in live soil from outside the rings. Further, seedling emergence in soil from inside the rings increased significantly in response to soil sterilisation. We found no significant difference in growth between sterile and live soils. However, this might be due to a lack of power caused by high rates of seedling mortality in all treatments. Overall, our study provides evidence for the role of soil pathogens in shaping this iconic Australian grass. Keywords: arid zone, Australia, culm, fire, hummock grassland, obligate seeder, seedling emergence, semi-arid zone, spinifex, soil pathogens, Triodia basedowii, clonal plants. Received 22 September 2020, accepted 23 February 2021, published online 29 March 2021 Introduction functionally similar to that of a shrub (Rice and Westoby Ring formation is a curiosity observed among many species of 1999; Nicholas et al. 2011), where large individuals can form clonal plants (Bonanomi et al. 2014; Watt 1947). Growth in domes up to 2 m high and more than 2 m in diameter (Burbidge ring-forming clonal plants is characterised by outward radial 1945; Lazarides 1997). Triodia hummocks form habitat and expansion and establishment of individual ramets followed by provide food for small mammals, lizards and birds (Murray progressive die-back of inner, older roots and shoots and Dickman 1994;Dalyet al. 2007;Brownet al. 2009)and (Bonanomi et al. 2005, 2014;Shefferet al. 2007; Cartenì accumulated biomass fuels cyclical wildfires initiating et al. 2012). Ring-forming species are often found in grassland landscape regeneration (Nicholas et al. 2009, 2011; Gamage communities (Bonanomi et al. 2014) and semi-arid or arid et al. 2012). Triodia species are valued by Australian regions (Sheffer et al. 2007; Ravi et al. 2008)including indigenous people, providing resins for use in traditional Australian hummock grasslands (Beadle 1981; Specht tool making and medicine as well as food and fibre 1981). In this study, we investigated ring formation in the (Gamage et al. 2012), and pastoralists use some species for Australian grass Triodia basedowii E.Pritz. We asked whether grazing (Lazarides 1997). As a result of their geographic soil microbes might contribute to the distinctive ring growth extent, Triodia grasses are of great ecological, cultural and form of T. basedowii which is commonly seen in Australia’s economic importance. arid regions. Ring formation occurs in several Triodia species (Burbidge The hummock grasslands of Australia’s arid and semi-arid 1945; Lazarides 1997). Of the known ring formers, Triodia interior are dominated by species in the genus Triodia R.Br. basedowii is widespread across the interior of Australia, (spinifex) and cover at least 1.3 Â 106 km2 or 18% of the growing among sand dunes from 19 to 30S (Lazarides continent (Griffin 1984; National Vegetation Information 1997;Grigget al. 2008). T. basedowii is killed by fire and System 2007). Although Triodia species are a grass, their is therefore an obligate seeder, relying on an accumulated soil ‘hummock’ growth form, consisting of a stiff tangle of culms seed bank for regeneration (Casson and Fox 1987;Westoby (hollow stems) and pungent leaves (Burbidge 1945), is et al. 1988; Rice and Westoby 1999). Hummocks comprise a Journal compilation Ó CSIRO 2021 Open Access CC BY-NC-ND www.publish.csiro.au/journals/ajb 114 Australian Journal of Botany N. D. Ross and A. T. Moles collection of individually rooted culms linked by stolons, (Bonanomi et al. 2005). Interestingly, the effects were species where the death of roots in older culms is thought to specific, with the same soils having much less effect on initiate die-back within the centre of ring (Burbidge 1945). seedlings of other species (Bonanomi et al. 2005). Models Westoby et al.(1988) found that recruitment in T. basedowii based on negative plant–soil feedbacks have successfully was concentrated along the outer edges of burnt hummocks recreated the real-world patterns seen in ring-forming despite similar densities of live seeds under hummocks and vegetation (Cartenì et al. 2012). around their edges. This outward bias in seedling recruitment One potential mechanism for ring formation that has not could explain ring-shaped hummocks in second-generation been studied in Triodia is the interaction of the plant with the plants, but does not explain death and collapse within older soil microbial community. Previous studies show that plants hummocks (Burbidge 1946). Thus, although outward radial can accumulate pathogens in their root zone over time (Bever growth with central die-back is an established phenomenon in 1994;VanderPuttenet al. 2013; Smith-Ramesh and Reynolds Triodia species (Burbidge 1945;Beard1984), little is known 2017). These pathogens can then weaken plant growth and of what causes the death of older culms. dominance, facilitating the coexistence of plant species within The fact that many ring-forming species occur in arid communities (Klironomos 2002; Bonanomi et al. 2012; ecosystems has led some researchers to suggest that water Reinhart 2012; Yang et al. 2015; Chung and Rudgers availability might drive ring-formation in some species 2016). In addition, soil pathogens can reduce interspecific (Getzin et al. 2016;Yizhaqet al. 2019;Herootyet al. competitive ability (Petermann et al. 2008;Hendrikset al. 2020). Although models and observational data are 2013) and shape succession of species within a community consistent with this idea in some ecosystems, other over time (Oremus and Otten 1981; Van der Putten et al. 1993; researchers have noted that ring-formation also occurs in Frouz et al. 2016). For example, seedlings grown in soil that several ecosystems in which water limitation is highly has been exposed to the roots of conspecifics have reduced unlikely to be important, including salt-marshes and alluvial biomass compared to seedlings grown in sterilised conspecific grasslands (Cartenì et al. 2012). Further, although the presence soil or soil exposed to other species (Packer and Clay 2003; of plants can affect soil moisture, the soil in the centre of Callaway et al. 2004; Gundale et al. 2014). Plants may Asphodelus ramosus rings in Israel have a comparable water temporarily escape their pathogens by colonising new soil content to the soil in the surrounding matrix (Yizhaq et al. through clonal growth (van der Stoel et al. 2002;Vander 2019). Thus, in this system, water availability does not seem to Putten 2003) or by dispersing offspring away from the parent provide a good explanation for ring-forming plants expanding plant (Packer and Clay 2000). In this way, the outward into the matrix while dying back inside rings. Getzin et al. expansion of ring-forming plants may be an escape (2019) also found that the texture and compaction (both factors response from accumulated soil pathogens, with die-back in that affect moisture absorption into soil) of soil inside Triodia the centre of the plant being due to older ramets being rings were not significantly different to that found in bare soil overwhelmed by soil pathogens. Consistent with this, a areas outside the rings. Finally, the proposed mechanisms study of Bouteloua gracilis in New Mexico, USA showed through which water availability might cause ring-formation that roots from the inner edge of grass rings had 1.4 times are not consistent across sites or species (Getzin et al. 2016; higher fungal colonisation in the field than did roots from Herooty et al. 2020). Experimental work on the role of water outside the rings (Carlton et al. 2018). While live soil had an availability in ring-formation would be a worthwhile next step, overall negative effect on plant growth, glasshouse studies did but this is not the goal of the present study. not show greater negative effects of soil from inside the rings Previous studies have rejected the idea that ants or termites (Carlton et al. 2018). Here, we aimed to test the idea that might be responsible for the die-back in the centre of Triodia pathogenic soil microbes might contribute to ring formation in hummocks (Getzin et al. 2016, 2019). Triodia. Specifically, we tested the hypothesis that microbes in Another possibility is that dieback in the centre of ring- the soil inside naturally formed T. basedowii rings negatively forming plants could be caused by depletion of soil nutrients. affect the emergence, survival and growth of T. basedowii Rice et al.(1994) found higher levels of soil nitrogen seedlings. concentrated within T. basedowii, T. pungens and Plectrachne schinzii hummocks compared to outside, Materials and methods whereas available soil phosphorous was lower inside hummocks compared to outside. However, the addition of Study site fertiliser or ash within the boundaries of hummocks did not We focused on Triodia basedowii growing naturally on flat, result in new growth over 4 months following summer rain interdune soil in the Northern Territory, Australia.